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Infection and Immunity, February 1999, p. 653-658, Vol. 67, No. 2
Division of AIDS,
Received 26 August 1998/Returned for modification 17
September 1998/Accepted 30 November 1998
A tissue culture bilayer system that mimics some aspects of early
alveolar infection by Mycobacterium tuberculosis was
developed. This model incorporates human lung epithelial type II
pneumocyte (A549) (upper chamber) and endothelial cell (lower chamber)
layers separated by a microporous membrane. This construction makes it possible to observe and quantify the passage of bacteria through the
two layers, to observe the interaction of the bacteria with the various
cell types, and to examine the basic mechanisms of immune cell
recruitment to the site of infection. After 107 organisms
were added to the upper chamber we microscopically observed large
numbers of bacteria attached to and within the pneumocytes and we
determined by viable-cell counting that a small percentage of the
inoculum (0.02 to 0.43%) passed through the bilayer into the lower
chamber. When peripheral blood mononuclear cells were added to the
lower chamber, microscopic examination indicated a migration of the
mononuclear cells through the bilayer to the apical surface, where they
were seen associated with the mycobacteria on the pneumocytes. The
added complexity of the bilayer system offers an opportunity to define
more precisely the roles of the various lung cell types in the
pathogenesis of early tuberculosis.
0019-9567/99/$00.00+0
An In Vitro Tissue Culture Bilayer Model To Examine
Early Events in Mycobacterium tuberculosis
Infection
*
Corresponding author. Mailing address: Bldg. 5, Rm.
B38, M/S G11, Centers for Disease Control and Prevention, Atlanta, GA 30333. Phone: (404) 639-3205. Fax: (404) 639-4192. E-mail:
fdq1{at}cdc.gov.
Infection and Immunity, February 1999, p. 653-658, Vol. 67, No. 2
0019-9567/99/$00.00+0
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